Lactobacillus gasseri 2459, isolated from the human vagina, exhibits a strong autoaggregating phenotype. Filter-sterilized spent supernatants of this strain promote aggregation of Lact. plantarum LL441 and Enterococcus faecalis EF. Aggregation was abolished upon exposure of the cells to proteases and, in the case of Ent. faecalis, to metaperiodate, which suggests the involvement of cell-surface proteins and glycoproteins, respectively, in the aggregation phenotype. In accordance with this, a 75 kDa surface protein, and possibly another of approximately 94 kDa, appears in Lact. plantarum LL441 cultures incubated with Lact. gasseri culture supernatants. The diffusible aggregation promoting factor was purified from stationary phase culture supernatants and determined to be a 2 kDa hydrophilic peptide active at pH 3-4 and stable at neutral and acid pH. The activity was resistant to heat, chymotrypsin, chelating agents, triton X-100 and reducing agents, but sensitive to other proteases and SDS.